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All-trans retinoic acid improves goat oocyte nuclear maturation and reduces apoptotic cumulus cells during in vitro maturation

机译:全反式维甲酸可改善山羊卵母细胞核成熟,并减少体外成熟过程中的凋亡积云细胞

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摘要

All-trans retinoic acid (t-RA) is a natural component and representative physiologically active metabolite of vitamin A, having multiple physiologic functions. The objective of this study was to evaluate the effect of t-RA on goat oocyte maturation and cumulus cell apoptosis during in vitro maturation (IVM). Immature goat cumulus-oocyte complexes (COCs) were matured in vitro in the absence or presence of t-RA at concentrations of 10nmol/L, 100nmol/L and 1000nmol/L. Oocyte maturation and embryo development were assessed by polar body formation and parthenogenetic activation, respectively. Oocyte survival was checked by Trypan blue staining. Apoptosis of cumulus cells was analyzed by terminal deoxynucleotidyl transferase nick end labeling staining and quantitative real-time PCR. In comparison with the control group, 100nmol/L and 10nmol/L t-RA significantly improved goat nuclear oocyte maturation and survival (P<0.05). Addition of 1000nmol/L t-RA improved nuclear maturation (P<0.05), but had no effect on survival of goat oocytes. t-RA had no positive effect on goat parthenogenetic embryonic cleavage, blastocyst formation or total cell numbers. However, t-RA inhibits the apoptosis of cumulus cells (P<0.01). t-RA treatment up-regulated the expression of B-cell lymphoma 2 (BCL-2), catalase (CAT) (P<0.05) and down-regulated the expression of Caspase-8 (P<0.05). In conclusion, t-RA has positive effects on goat oocyte nuclear maturation and reduces apoptotic cumulus cells during IVM.
机译:全反式维甲酸(t-RA)是维生素A的天然成分和代表生理活性代谢产物,具有多种生理功能。这项研究的目的是评估t-RA对体外成熟(IVM)期间山羊卵母细胞成熟和卵丘细胞凋亡的影响。在不存在或存在t-RA的情况下,浓度为10nmol / L,100nmol / L和1000nmol / L的未成熟山羊卵母细胞复合物(COC)在体外成熟。卵母细胞成熟和胚胎发育分别通过极体形成和孤雌生殖活化来评估。通过台盼蓝染色检查卵母细胞存活。通过末端脱氧核苷酸转移酶切口末端标记染色和定量实时PCR分析卵丘细胞的凋亡。与对照组相比,100nmol / L和10nmol / L的t-RA显着改善了山羊核卵母细胞的成熟和存活率(P <0.05)。添加1000nmol / L t-RA可以促进核成熟(P <0.05),但对山羊卵母细胞的存活没有影响。 t-RA对山羊孤雌胚胎分裂,胚泡形成或总细胞数无积极影响。然而,t-RA抑制卵丘细胞的凋亡(P <0.01)。 t-RA治疗上调B细胞淋巴瘤2(BCL-2),过氧化氢酶(CAT)的表达(P <0.05),下调Caspase-8的表达(P <0.05)。总之,t-RA对山羊卵母细胞核成熟具有积极作用,并在IVM期间减少凋亡性丘疹细胞。

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